Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode
Abstract Plant parasitic nematodes (PPNs) cause significant damage to crop production worldwide, leading to substantial economic losses. Conventional chemical nematicides are effective but frequently associated with environmental and health hazards. In response, biological control methods, particula...
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SpringerOpen
2024-12-01
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| Series: | Chemical and Biological Technologies in Agriculture |
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| Online Access: | https://doi.org/10.1186/s40538-024-00720-8 |
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| author | Peiyu Yan Shakil Ahmad Zhixia Xu He Jia Renyue Zhang Jingwen Song Nazia Manzar Abhijeet Shankar Kashyap Wenfei Zhang |
| author_facet | Peiyu Yan Shakil Ahmad Zhixia Xu He Jia Renyue Zhang Jingwen Song Nazia Manzar Abhijeet Shankar Kashyap Wenfei Zhang |
| author_sort | Peiyu Yan |
| collection | DOAJ |
| description | Abstract Plant parasitic nematodes (PPNs) cause significant damage to crop production worldwide, leading to substantial economic losses. Conventional chemical nematicides are effective but frequently associated with environmental and health hazards. In response, biological control methods, particularly the use of microbial pesticides, have emerged as a sustainable and effective alternative. This study focuses on the isolation and characterization of Bacillus sp. HSY32, a bacterial strain with nematicidal properties, from a tropical rainforest soil sample in Hainan, China. Soil samples were screened for nematicidal activity, which led to the identification of the strain HSY32. Detailed observations using optical and scanning electron microscopy (SEM) revealed that HSY32 forms spores and parasporal crystal structures, which are typically associated with nematicidal Bacillus species. Genomic analysis of HSY32 showed that its genome spans 6,711,949 base pairs and contains 7915 predicted genes, with an average GC content of 35.4%. Phylogenetic analysis, utilizing 16S rRNA sequences and average nucleotide identity (ANI), established that HSY32 is closely related to Bacillus mobilis, a known species within the Bacillus genus. Further genomic analysis using local BLAST identified several toxin genes with high similarity to known nematicidal genes, including cry4Ba, cry50Ba, app6Ba, cry70Bb, and tpp36Aa. To confirm the functionality of these toxin genes, they were cloned into pET-30a expression vectors and expressed in E. coli BL21 (DE3) cells. Among the expressed proteins, the Cry4Ba-like protein, with a molecular weight of approximately 110 kDa, was found to exhibit significant nematicidal activity in bioassays. This protein demonstrated the ability to kill or inhibit the growth of PPNs, indicating its potential as a biological control agent. The successful isolation of Bacillus strain HSY32 and the identification of its novel Cry4-like toxin gene represent a significant advancement in the field of biological control of plant parasitic nematodes. The nematicidal activity of the Cry4Ba-like protein highlights the potential of HSY32 as a source of new biopesticides. Further studies are required to enhance the production and application of these biocontrol agents in agriculture, paving the way for more sustainable and eco-friendly methods to control PPNs. Graphical Abstract |
| format | Article |
| id | doaj-art-033394532c2a4271ba7fbbf306f37312 |
| institution | DOAJ |
| issn | 2196-5641 |
| language | English |
| publishDate | 2024-12-01 |
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| series | Chemical and Biological Technologies in Agriculture |
| spelling | doaj-art-033394532c2a4271ba7fbbf306f373122025-08-20T02:57:30ZengSpringerOpenChemical and Biological Technologies in Agriculture2196-56412024-12-0111111210.1186/s40538-024-00720-8Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematodePeiyu Yan0Shakil Ahmad1Zhixia Xu2He Jia3Renyue Zhang4Jingwen Song5Nazia Manzar6Abhijeet Shankar Kashyap7Wenfei Zhang8Ministry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityPlant Pathology Lab, ICAR-National Bureau of Agriculturally Important MicroorganismsPlant Pathology Lab, ICAR-National Bureau of Agriculturally Important MicroorganismsMinistry of Education Key Laboratory for Ecology of Tropical Islands, College of Life Sciences, Hainan Normal UniversityAbstract Plant parasitic nematodes (PPNs) cause significant damage to crop production worldwide, leading to substantial economic losses. Conventional chemical nematicides are effective but frequently associated with environmental and health hazards. In response, biological control methods, particularly the use of microbial pesticides, have emerged as a sustainable and effective alternative. This study focuses on the isolation and characterization of Bacillus sp. HSY32, a bacterial strain with nematicidal properties, from a tropical rainforest soil sample in Hainan, China. Soil samples were screened for nematicidal activity, which led to the identification of the strain HSY32. Detailed observations using optical and scanning electron microscopy (SEM) revealed that HSY32 forms spores and parasporal crystal structures, which are typically associated with nematicidal Bacillus species. Genomic analysis of HSY32 showed that its genome spans 6,711,949 base pairs and contains 7915 predicted genes, with an average GC content of 35.4%. Phylogenetic analysis, utilizing 16S rRNA sequences and average nucleotide identity (ANI), established that HSY32 is closely related to Bacillus mobilis, a known species within the Bacillus genus. Further genomic analysis using local BLAST identified several toxin genes with high similarity to known nematicidal genes, including cry4Ba, cry50Ba, app6Ba, cry70Bb, and tpp36Aa. To confirm the functionality of these toxin genes, they were cloned into pET-30a expression vectors and expressed in E. coli BL21 (DE3) cells. Among the expressed proteins, the Cry4Ba-like protein, with a molecular weight of approximately 110 kDa, was found to exhibit significant nematicidal activity in bioassays. This protein demonstrated the ability to kill or inhibit the growth of PPNs, indicating its potential as a biological control agent. The successful isolation of Bacillus strain HSY32 and the identification of its novel Cry4-like toxin gene represent a significant advancement in the field of biological control of plant parasitic nematodes. The nematicidal activity of the Cry4Ba-like protein highlights the potential of HSY32 as a source of new biopesticides. Further studies are required to enhance the production and application of these biocontrol agents in agriculture, paving the way for more sustainable and eco-friendly methods to control PPNs. Graphical Abstracthttps://doi.org/10.1186/s40538-024-00720-8Biological controlBacillus sp. HSY32Cry4-like toxinPlant parasitic nematodesMicrobial pesticides |
| spellingShingle | Peiyu Yan Shakil Ahmad Zhixia Xu He Jia Renyue Zhang Jingwen Song Nazia Manzar Abhijeet Shankar Kashyap Wenfei Zhang Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode Chemical and Biological Technologies in Agriculture Biological control Bacillus sp. HSY32 Cry4-like toxin Plant parasitic nematodes Microbial pesticides |
| title | Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode |
| title_full | Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode |
| title_fullStr | Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode |
| title_full_unstemmed | Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode |
| title_short | Isolation and characterization of Bacillus sp. HSY32 and its toxin gene for potential biological control of plant parasitic nematode |
| title_sort | isolation and characterization of bacillus sp hsy32 and its toxin gene for potential biological control of plant parasitic nematode |
| topic | Biological control Bacillus sp. HSY32 Cry4-like toxin Plant parasitic nematodes Microbial pesticides |
| url | https://doi.org/10.1186/s40538-024-00720-8 |
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